含N双杂环酰胺类化合物的设计、合成及免疫抑制活性的初步研究

发布时间：2018-03-13 16:37

本文选题：PI3K/mTOR　切入点：免疫抑制剂　出处：《重庆理工大学》2014年硕士论文　论文类型：学位论文

【摘要】：免疫抑制剂(immunosuppressant)是一类通过抑制细胞及体液免疫反应而使组织损伤得以减轻的物质。它可抑制机体异常的免疫反应，主要用来治疗器官移植抗排斥反应和自身免疫性疾病。虽然近年来，新型免疫抑制剂不断问世，但多数具有选择性不高，可降低机体的正常免疫应答、诱发感染和肿瘤等问题。研究发现PI3K/Akt/mTOR信号通路在一些肿瘤及免疫性疾病中处于激活状态。虽然已经研发出专门针对单一蛋白激酶的免疫抑制剂。但是这些抑制剂在治疗免疫疾病时仍然存在一些弊端。为了寻找新型选择性强、作用快速、效果明显、不良反应少的免疫抑制剂，本课题在计算机辅助药物设计（CADD）的基础上，以PI3K蛋白激酶和mTOR蛋白为受体，筛选出先导化合物，，并运用逆合成的方法，合成了一系列具有免疫抑制活性的含N双杂环酰胺类化合物。为进一步开发新型的PI3K/mTOR双重免疫抑制剂提供了实验依据。本课题研究的主要内容： 1）先导化合物的确定运用Sybyl2.0分别以PI3K和mTOR作为受体，对ZINC,PubMed和BindingDB中下载到的近200万个小分子进行虚拟筛选。通过对虚筛得到的小分子进行生物活性测试，确定先导化合物。 2）含N双杂环酰胺类化合物的合成及纯化运用逆合成的方法，分析合成路线，以最合理的合成方案合成一系列先导化合物的衍生物。通过液质联用技术以及波普技术，分析确定其结构。 3）含N双杂环酰胺类化合物对Jurkat T淋巴细胞的活性测定基于CCK-8技术，测定所合成的化合物对于T淋巴细胞系的急性白血病细胞Jurkat的体外抑制活性。通过上述研究，所得研究成果如下： 1）确定了3-(2,6-二氯苯基)-5-甲基-N-2-(6-甲磺酰苯并噻唑基)异恶唑-4-甲酰胺作为先导化合物。在此基础上以2,6-二氯苯甲醛为起始原料经过6步反应，合成了包含先导化合物在内的12个目标化合物。经高效液相（HPLC）检测纯度都达到98%以上，质谱数据与核磁数据均与目标化合物相符。 2）经CCK-8法测试含N双杂环酰胺类化合物对Jurkat细胞的增值抑制，获得了有效可靠的数据。验证了化合物对于Jurkat T淋巴细胞的杀伤作用。 3)本课题所设计的含氮双杂环酰胺类化合物具有良好的免疫抑制作用，为进一步研究该类化合物，开发PI3K/mTOR双重抑制剂提供了良好的实验基础。
[Abstract]:Immunosuppressant, an immunosuppressant, is a class of substances that attenuate tissue damage by inhibiting cellular and humoral immune responses. Although new immunosuppressants have been developed in recent years, most of them have low selectivity, which can reduce the normal immune response, induce infection and tumor. Studies have found that PI3K/Akt/mTOR signaling pathways are activated in some tumors and immune diseases. Although immunosuppressants have been developed specifically for single protein kinases, these inhibitors are still present in the treatment of immune diseases. There are some disadvantages. In order to find a new type of selectivity, Immunosuppressants with rapid effect, obvious effect and less adverse reactions. Based on the computer aided drug design (CADDD), PI3K protein kinase and mTOR protein were used as receptors to screen the leading compounds, and the reverse synthesis method was used. A series of N-containing bicyclic amides with immunosuppressive activity were synthesized, which provide experimental basis for the further development of novel PI3K/mTOR double immunosuppressants. The main contents of this research are as follows:. 1) determination of lead compounds. PI3K and mTOR were used as receptors for virtual screening of nearly 2 million small molecules downloaded from BindingDB and pubMed by Sybyl2.0, and the leading compounds were determined by bioactivity test of the small molecules obtained by virtual screening. 2) Synthesis and purification of N-containing bicyclic amides. A series of derivatives of leading compounds were synthesized by the method of inverse synthesis, and their structures were determined by liquid-mass spectrometry and pop technique. 3) determination of the activity of N-containing bicyclic amides on Jurkat T lymphocytes. Based on CCK-8 technique, the inhibitory activity of the synthesized compounds on Jurkat of T lymphocyte line acute leukemia cells in vitro was determined. The results of the above studies are as follows:. 1) 3-trichlorobenzene 6-dichlorobenzene (2-Dichlorobenzyl) -5-methyl-N-2-thiazolyl) isoxazole-4-formamide was determined as the lead compound. On the basis of this, the starting material of 2C6-dichlorobenzaldehyde was used as the starting material, and the reaction was carried out in six steps. Twelve target compounds, including lead compounds, were synthesized. The purity of these compounds was over 98% by HPLC, and the mass spectrum data and the NMR data were all consistent with the target compounds. 2) the inhibitory effect of N-containing bicyclic amides on the proliferation of Jurkat cells was tested by CCK-8 method, and the effective and reliable data were obtained, and the killing effect of the compounds on Jurkat T lymphocytes was verified. 3) the nitrogen-containing bicyclic amides have good immunosuppressive effect, which provides a good experimental basis for the further study of these compounds and the development of PI3K/mTOR double inhibitors.
【学位授予单位】：重庆理工大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R914.5;R96

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